Microfluidic protein isolation and sample preparation for high-resolution cryo-EM

• DOI: 10.1073/pnas.1907214116
• Publication Date: 2019-07-10T23:27:17Z
• Authors: Claudio Schmidli; Stefan Albiez; Luca Rima; Ricardo Righetto; Inayatulla Mohammed; Paolo Oliva; Lubomir Kovacik; Henning Stahlberg; Thomas Braun
• Keywords: Cryo-Electron Microscopy; Sample Preparation; Target protein; Isolation

High-resolution structural information is essential to understand protein function. Protein-structure determination needs a considerable amount of protein, which can be challenging produce, often involving harsh and lengthy procedures. In contrast, the several thousand few million particles required for structure by cryogenic electron microscopy (cryo-EM) provided miniaturized systems. Here, we present microfluidic method rapid isolation target its direct preparation cryo-EM. Less than 1 μL cell lysate as starting material solve atomic untagged, endogenous human 20S proteasome. Our work paves way high-throughput proteins from minimal amounts opens more opportunities sensitive, complexes.